A thermosensitive polymer coupled to nucleic acid or protein and its application
A technology for coupling nucleic acids and polymers, which is applied in the field of temperature-sensitive polymer materials, can solve the problems of poor versatility of temperature-sensitive materials, and achieve the effect of low production cost, low price, and cheap and easy-to-obtain raw materials
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Embodiment 1
[0040] 1. Use a pipette gun to take 100 μL streptavidin (2×10 -3 g / mL) and 250 μL coupling agent N-hydroxysuccinimide acrylate (1×10 -3 g / mL) in a centrifuge tube, mix evenly, transfer to an incubator, shake and react at 37°C for 1 hour to obtain a coupling product;
[0041] 2. Add 0.06g of N-isopropylacrylamide monomer and 4650μL of PBS buffer solution (NaCl concentration: 15nM) to the centrifuge tube in turn, mix with a vortex mixer until the N-isopropylacrylamide monomer is completely dissolved, Obtain a mixed solution, then add 10 mg of ammonium persulfate and 5 μL of tetramethylethylenediamine to the mixed solution in sequence, mix evenly, move to an incubator, react and polymerize at 25°C for 2 hours, and obtain a compound containing coupled streptavidin Mixed systems of thermosensitive polymers;
[0042]3. Move the mixed system containing the temperature-sensitive polymer coupled with streptavidin to a water bath, and incubate at 37°C for 5 minutes, so that the phase ...
Embodiment 2
[0108] 1. Use a pipette gun to take 250 μL of nucleic acid sequences modified with NH2 at the 5' end and biotin at the 3' end (NH 2 -GGGTTGGGCGGGATGGGTTT-Biotin) (1×10 -6 mol / L) and 50 μL coupling agent N-hydroxysuccinimide acrylate (1×10 -3 g / mL) in a centrifuge tube, mixed evenly, transferred to an incubator, and reacted with shaking at 37°C for 1 hour to obtain a coupling product;
[0109] 2. Add 0.02g of N-isopropylacrylamide monomer and 700μL of PBS buffer solution (NaCl concentration: 15nM) to the centrifuge tube in turn, mix with a vortex mixer until the N-isopropylacrylamide monomer is completely dissolved, Obtain a mixed solution, then add 2 mg of ammonium persulfate and 1 μL of tetramethylethylenediamine to the mixed solution in sequence, mix well, move to an incubator, react and polymerize at 25°C for 2 hours, and obtain a thermosensitive compound containing coupled nucleic acid. Mixed systems of polymers;
[0110] 3. Move the mixed system to a water bath and kee...
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